Synthesis and reactivity of aryl nitrogen mustard-oligodeoxyribonucleotide conjugates.
A versatile method is described for preparing aryl nitrogen mustard-oligodeoxyribonucleotide (mustard-ODN) conjugates under anhydrous conditions. The chemistry uses DMSO soluble triethylammonium or tributylammonium salts of the ODNs. A G/A motif triplex forming ODN was chosen for study since it had been shown earlier to bind with high affinity and specificity to a duplex DNA target. A 5'-hexylamine derivative of this ODN was reacted with three different 2,3,5,6-tetrafluorophenyl ester derivatives of aryl nitrogen mustards which were designed to have different alkylation rates. An HPLC assay was used to determine reaction rates of these mustard-ODNs under various conditions. The reactivity of the mustard groups depended on chloride concentration and the presence of nucleophiles. Conjugation of mustards to G/A-containing ODNs decreased their aqueous stability. Hydrolysis and alkylation rates of these agents were consistent with reaction via an aziridinium intermediate. Rates of sequence specific alkylation within a triplex were determined by denaturing gel electrophoresis and shown to depend on inherent reactivity of the mustard group. The improved synthesis and chemical characterization of mustard-ODNs should facilitate their use as sequence specific alkylating agents and as probes for nucleic acid structure.
Reed
M W
MW
Epoch Pharmaceuticals, Inc., Bothell, Washington 98021, USA.
Lukhtanov
E A
EA
Gorn
V
V
Kutyavin
I
I
Gall
A
A
Wald
A
A
Meyer
R B
RB
D000478:Alkylation; D001483:Base Sequence; D002021:Buffers; D002851:Chromatography, High Pressure Liquid; D004247:DNA; D004121:Dimethyl Sulfoxide; D004355:Drug Stability; D006868:Hydrolysis; D007700:Kinetics; D009588:Nitrogen Mustard Compounds / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D009696:Nucleic Acids / Q000737:chemistry; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012995:Solubility
D002021:Buffers; D009588:Nitrogen Mustard Compounds; D009696:Nucleic Acids; D009838:Oligodeoxyribonucleotides; D000644:Quaternary Ammonium Compounds; C051802:triplex DNA; D004247:DNA; D004121:Dimethyl Sulfoxide